Development of Distributed Feedback Laser Biosensor Technology as a High Resoluti

描述(由申请人提供)：该提案描述了一种全新类型的光学生物传感器的初步开发，该传感器利用基于塑料的激光表面结构的主动光输出来提供对生物分子相互作用的高分辨率无标记传感。虽然激光生物传感器可以通过复制模塑工艺在较大的表面积上廉价地生产，但我们建议建立的检测仪器将是紧凑、廉价的，并且能够分辨率足以直接检测与高信噪比的固定蛋白质靶标结合的小分子。这里提出的第一项资助工作将使传感器制造工艺的开发、检测仪器的建造以及一系列小分子直接结合分析的初步可行性研究得以进行。我们的长期目标是将这种生物传感器方法发展成为一个通用的检测平台，利用其在成像检测和单蛋白质分子检测方面的潜力，用于小分子筛选、细胞分析、生物标记物诊断和环境检测。 与公共卫生相关：拟议的项目旨在开发一种高分辨率的生物分子相互作用检测技术，该技术可用于处理由不具有酶功能的蛋白质组成的大片蛋白质组，因此不能使用传统技术进行“筛选”。调控转录的蛋白质、从细胞表面接收信号的蛋白质，以及参与DNA损伤修复的蛋白质，都在数千种蛋白质中，这些蛋白质不具有酶活性，但它们通过药物的调节可能会导致治疗一系列疾病。新的蛋白质靶点的数量停滞不前，突显了对高通量筛选新技术的需求：在1989-2000年的几年里，只有6%的新分子实体击中了以前未被下药的靶点。因此，为了找到新的靶点并确定在疾病治疗中真正取得突破的新化合物，我们需要能够用来快速识别蛋白质的小分子结合物的检测方法，而不仅仅是酶抑制剂。

DESCRIPTION (provided by applicant): The proposal describes the initial development of a fundamentally new type of optical biosensor that utilizes active generation of light output with a plastic-based laser surface structure to provide high resolution label-free sensing of biomolecular interactions. While the laser biosensor can be produced inexpensively over large surface areas by a replica molding process, the detection instrument that we propose building will be compact, inexpensive, and capable of resolution sufficient for direct detection of small molecule binding to immobilized protein targets with high signal-to-noise ratio. The first funded effort, proposed here, would enable development of the sensor fabrication process, construction of a detection instrument, and performance of a set of initial feasibility studies for small molecule direct binding assays. Our long-term goal is to develop this biosensor method into a general-purpose detection platform for small molecule screening, cell-based assays, biomarker diagnostics, and environmental detection by taking advantage of its potential for imaging detection and single protein molecule detection resolution. PUBLIC HEALTH RELEVANCE: The proposed project is aimed at the development of a high resolution biomolecular interaction detection technology that can be utilized to address the large swath of the proteome consisting of proteins that have no enzymatic function and are hence not "screenable" using conventional technologies. Proteins that modulate transcription, those that receive signals from the cellular surface, and those that are involved in DNA damage repair are among the thousands of proteins that do not have enzymatic activity but whose modulation with drugs could lead to treatments for a host of diseases. The stagnation in the number of new protein targets underscores the need for new technologies for high-throughput screening: in the years from 1989-2000, only 6% of the new molecular entities hit previously undrugged targets. Thus, to get at new targets and identify new compounds that are true breakthroughs in disease treatment, we need detection methodologies that can be utilized to rapidly identify small molecule binders for proteins, not merely enzymatic inhibitors.